Coronary disease testing method and reagent kit
A kit and in vitro detection technology, applied in the fields of molecular biology and medicine, can solve the unconfirmed correlation between F11R gene and atherosclerotic coronary heart disease, unconfirmed F11R gene SNP and atherosclerosis and/or coronary heart disease issues of relevance
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0068] 1.1 Research object
[0069] Among the patients diagnosed with atherosclerosis by coronary angiography, the patients with 2 or more lesions and a stenosis of more than 50% were selected as cases. In addition, normal people without atherosclerosis diagnosed by coronary angiography were selected as controls, and the age, sex, and place of origin were matched with the case group as much as possible.
[0070] Peripheral blood samples were randomly collected from 191 patients and 186 normal controls on the basis of informed consent.
[0071] 1.2 Experimental methods and results
[0072] 1.2.1 DNA extraction
[0073] DNA was extracted from human peripheral blood samples by the conventional phenol-chloroform method, and the concentration was corrected to 20ng / ul for conventional PCR amplification.
[0074] 1.2.2 Design of PCR and sequencing primers
[0075] According to the genome sequence of F11R in GenBank, the following primers were designed and synthesized. The specif...
Embodiment 2
[0089] Atherosclerosis and / or Coronary Heart Disease Susceptibility Detection Kit
[0090] As described in Example 1, the T→C mutation at position 61 in SEQ ID NO: 1 is closely related to atherosclerosis and / or coronary heart disease. Therefore, specific primers for the F11R gene can be designed based on this mutation and then amplified using the patient's DNA as a template for detection.
[0091] Prepare a test kit (100 person-times), which contains:
[0092]
[0093] A test group composed of 100 people was randomly selected, including subjects who were unknown whether they had atherosclerosis, patients who were known to have atherosclerosis and / or coronary heart disease, and normal people who had been tested without atherosclerosis.
[0094] Take 3ml of peripheral blood from the subject to be tested in the test group, and use conventional methods (or use a specific kit) to extract DNA from the blood. Dilute the PCR primers in the atherosclerosis and / or coronary heart di...
Embodiment 3
[0100] Auxiliary testing for susceptibility to atherosclerosis and / or coronary heart disease
[0101] The test in Example 2 was repeated, with the difference that 80 people (whose symptoms of atherosclerosis were not known before the test) were randomly selected for testing.
[0102] Prepare a test kit (100 person-times), which contains:
[0103]
[0104] Take 3ml of peripheral blood from the subject to be tested, and use conventional methods (or use a specific kit) to extract DNA from the blood. Dilute the PCR primers in the atherosclerosis and / or coronary heart disease detection kit to 1μmol / μl, and use the extracted DNA as a template to perform a PCR reaction with the provided primers. After purification of PCR products, ABI-PRISM TM 377 DNA sequencer was used for two-way sequencing by fluorescence-labeled terminal termination method, and Polyphred software was used for sequence interpretation and SNP confirmation.
[0105] The results also confirmed that the proport...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More